Assessment of guanylin peptides system in type 2 diabetes

Os doentes com diabetes mellitus tipo 2 apresentam predisposição para a retenção de sódio e são frequentemente hipertensos. No entanto, os mecanismos implicados na dificuldade do rim diabético em mobilizar o sódio são, ainda, pouco compreendidos. Os peptídeos da família das guanilinas estão envolvidos na regulação do transporte de electrólitos e água nos epitélios intestinal e renal, através da activação do receptor guanilato ciclase-C (GC-C) e subsequente libertação intracelular de GMPc. O objectivo do presente estudo foi a avaliação da actividade do sistema dos peptídeos das guanilinas (SPG) e do seu papel na regulação do balanço de sódio num modelo animal de diabetes tipo 2. Ratinhos machos C57BL/6 foram submetidos a uma dieta com alto teor de gordura e rica em hidratos de carbono simples (ratinhos diabéticos) ou a uma dieta normal (ratinhos controlo). A expressão renal e intestinal da guanilina (GN), uroguanilina (UGN) e do receptor GC-C assim como os níveis de GMPc na urina e plasma foram avaliados nos ratinhos controlo e diabéticos, durante a ingestão de dietas normo (NS) e hiper-salina (HS). Nos ratinhos diabéticos, durante a dieta NS verificou-se um aumento significativo da pressão arterial que foi acompanhado de redução da expressão do ARNm da GN, UGN e do GC-C no intestino e de aumento da expressão de ARNm da UGN no rim. A dieta HS induziu um aumento da expressão do ARNm da UGN no jejuno dos ratinhos controlo mas não nos diabéticos. Os ratinhos diabéticos apresentaram níveis urinários de GMPc inferiores aos controlos, em condições de dieta NS. Em conclusão, os nossos resultados sugerem que na diabetes tipo 2 ocorre uma redução da actividade intestinal do SPG que é acompanhada por um aumento compensatório da actividade renal do SPG. A diminuição da actividade do SPG intestinal na diabetes tipo 2 deve-se não só a uma redução da expressão dos peptídeos GN e UGN, mas também a uma redução da expressão do seu receptor, GC-C. Estes resultados sugerem que o SPG pode contribuir para a sensibilidade ao sódio na diabetes.

Bradykinin-induced Ca2+ signaling in human subcutaneous fibroblasts involves ATP release via hemichannels leading to P2Y12 receptors activation

Background: Chronic musculoskeletal pain involves connective tissue remodeling triggered by inflammatory mediators, such as bradykinin. Fibroblast cells signaling involve changes in intracellular Ca2+ ([Ca2+]i). ATP has been related to connective tissue mechanotransduction, remodeling and chronic inflammatory pain, via P2 purinoceptors activation. Here, we investigated the involvement of ATP in bradykinin-induced Ca2+ signals in human subcutaneous fibroblasts. Results: Bradykinin, via B2 receptors, caused an abrupt rise in [Ca2+]i to a peak that declined to a plateau, which concentration remained constant until washout. The plateau phase was absent in Ca2+-free medium; [Ca2+]i signal was substantially reduced after depleting intracellular Ca2+ stores with thapsigargin. Extracellular ATP inactivation with apyrase decreased the [Ca2+]i plateau. Human subcutaneous fibroblasts respond to bradykinin by releasing ATP via connexin and pannexin hemichannels, since blockade of connexins, with 2- octanol or carbenoxolone, and pannexin-1, with 10Panx, attenuated bradykinin-induced [Ca2+]i plateau, whereas inhibitors of vesicular exocytosis, such as brefeldin A and bafilomycin A1, were inactive. The kinetics of extracellular ATP catabolism favors ADP accumulation in human fibroblast cultures. Inhibition of ectonucleotidase activity and, thus, ADP formation from released ATP with POM-1 or by Mg2+ removal from media reduced bradykinin-induced [Ca2+]i plateau. Selective blockade of the ADP-sensitive P2Y12 receptor with AR-C66096 attenuated bradykinin [Ca2+]i plateau, whereas the P2Y1 and P2Y13 receptor antagonists, respectively MRS 2179 and MRS 2211, were inactive. Human fibroblasts exhibited immunoreactivity against connexin-43, pannexin-1 and P2Y12 receptor. Conclusions: Bradykinin induces ATP release from human subcutaneous fibroblasts via connexin and pannexin-1-containing hemichannels leading to [Ca2+]i mobilization through the cooperation of B2 and P2Y12 receptors.

Characterization and Biological Activities of Ocellatin Peptides from the Skin Secretion of the Frog Leptodactylus pustulatus

Eight new peptides were isolated from the skin secretion of the frog Leptodactylus pustulatus and their amino acid sequences determined by de novo sequencing and by cDNA cloning. Structural similarities between them and other antimicrobial peptides from the skin secretion of Leptodactylus genus frogs were found. Ocellatins-PT1 to -PT5 (25 amino acid residues) are amidated at the C-terminus, while ocellatins-PT6 to -PT8 (32 amino acid residues) have free carboxylates. Antimicrobial activity, hemolytic tests, and cytotoxicity against a murine fibroblast cell line were investigated. All peptides, except for ocellatin-PT2, have antimicrobial activity against at least one Gram negative strain. Ocellatin-PT8 inhibited the growth of Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Salmonella choleraesuis strains with MICs in the 60−240 μM range. No significant effect was observed in human erythrocytes and in a murine fibroblast cell line after exposure to the peptides at MICs. A comparison between sequences obtained by both direct HPLC-MS de novo sequencing and cDNA cloning demonstrates the secretion of mature peptides derived from a pre-pro-peptide structure.

Are known cyanotoxins involved in the toxicity of picoplanktonic and filamentous north atlantic marine cyanobacteria?

Eight marine cyanobacteria strains of the genera Cyanobium, Leptolyngbya, Oscillatoria, Phormidium, and Synechococcus were isolated from rocky beaches along the Atlantic Portuguese central coast and tested for ecotoxicity. Strains were identified by morphological characteristics and by the amplification and sequentiation of the 16S rDNA. Bioactivity of dichloromethane, methanol and aqueous extracts was assessed by the Artemia salina bioassay. Peptide toxin production was screened by matrix assisted laser desorption/ionization time of flight mass spectrometry. Molecular analysis of the genes involved in the production of known cyanotoxins such as microcystins, nodularins and cylindrospermopsin was also performed. Strains were toxic to the brine shrimp A. salina nauplii with aqueous extracts being more toxic than the organic ones. Although mass spectrometry analysis did not reveal the production of microcystins or other known toxic peptides, a positive result for the presence of mcyE gene was found in one Leptolyngbya strain and one Oscillatoria strain. The extensive brine shrimp mortality points to the involvement of other unknown toxins, and the presence of a fragment of genes involved in the cyanotoxin production highlight the potential risk of cyanobacteria occurrence on the Atlantic coast.

An electrochemical deamidated gliadin antibody immunosensor for celiac disease clinical diagnosis

The first electrochemical immunosensor (EI) for the detection of antibodies against deamidated gliadin peptides (DGP) is described here. A disposable nanohybrid screen-printed carbon electrode modified with DGP was employed as the transducer's sensing surface. Real serumsampleswere successfully assayed and the results were corroborated with an ELISA kit. The presented EI is a promising analytical tool for celiac disease diagnosis.

Intelligent Micro Grid Management using a Multi-Agent approach

Recent changes in electricity markets (EMs) have been potentiating the globalization of distributed generation. With distributed generation the number of players acting in the EMs and connected to the main grid has grown, increasing the market complexity. Multi-agent simulation arises as an interesting way of analysing players’ behaviour and interactions, namely coalitions of players, as well as their effects on the market. MASCEM was developed to allow studying the market operation of several different players and MASGriP is being developed to allow the simulation of the micro and smart grid concepts in very different scenarios This paper presents a methodology based on artificial intelligence techniques (AI) for the management of a micro grid. The use of fuzzy logic is proposed for the analysis of the agent consumption elasticity, while a case based reasoning, used to predict agents’ reaction to price changes, is an interesting tool for the micro grid operator.

Load Control Timescales Simulation in a Multi-Agent Smart Grid Platform

Environmental concerns and the shortage in the fossil fuel reserves have been potentiating the growth and globalization of distributed generation. Another resource that has been increasing its importance is the demand response, which is used to change consumers’ consumption profile, helping to reduce peak demand. Aiming to support small players’ participation in demand response events, the Curtailment Service Provider emerged. This player works as an aggregator for demand response events. The control of small and medium players which act in smart grid and micro grid environments is enhanced with a multi-agent system with artificial intelligence techniques – the MASGriP (Multi-Agent Smart Grid Platform). Using strategic behaviours in each player, this system simulates the profile of real players by using software agents. This paper shows the importance of modeling these behaviours for studying this type of scenarios. A case study with three examples shows the differences between each player and the best behaviour in order to achieve the higher profit in each situation.

Autoavaliação das organizações escolares: articulação da equipa de autoavaliação com as lideranças intermédias

O presente trabalho de projeto resulta de um estudo na área da Avaliação Institucional, realizado num agrupamento de escolas com educação pré-escolar e ensinos básico e secundário. O objetivo principal relacionou-se com a construção de um Plano de Ação, que promova o desenvolvimento de uma cultura de avaliação, procurando envolver todos os ‘atores’ educativos no processo de mudança e melhoria do agrupamento, com principal incidência na articulação da Equipa de Autoavaliação do Agrupamento, com as Lideranças Intermédias, de forma a implementar o processo de Autoavaliação do Agrupamento. A avaliação das escolas deve basear-se em dispositivos simples e exequíveis e nesse sentido é importante que a avaliação respeite critérios de pertinência, de coerência, de eficácia, de eficiência e de oportunidade. As Equipas de Autoavaliação, em articulação com as Lideranças Intermédias, assumem cada vez mais importância, nas organizações escolares, podendo ser potenciadoras da melhoria da eficácia e eficiência das organizações escolares. Um dos instrumentos de recolha de dados foi o inquérito por questionário, aplicado a 89 professores e educadores, que representam 51,2 % dos docentes do agrupamento em estudo. Outro instrumento de recolha de dados foi o focus group, constituído por 11 professores, dos departamentos curriculares existentes no agrupamento. Os resultados mostraram que o processo de autoavaliação é já uma prática comum no agrupamento, com uma certa consolidação. No entanto, ao nível da estrutura que lidera, apesar de integrar representantes de toda a comunidade educativa, ainda se verifica uma participação muito centrada nos docentes. Esta estrutura deve tornar-se mais abrangente, de forma a permitir a incorporação de outros pontos de vista. Os resultados mostraram ainda, diferentes tipos de abordagem aos assuntos relacionados com a autoavaliação, nos distintos departamentos curriculares. O incentivo das Lideranças Intermédias e a sua implicação na tarefa de motivar os professores para a participação no processo, a articulação com a Equipa de Autoavaliação e a comunicação serão determinantes para a melhoria da eficácia e eficiência da organização. No plano de ação, é apresentado um conjunto de propostas, que pretendem colmatar estas debilidades.